| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
* Department of Biomedical Engineering,
Cardiovascular Research Center, and
Center for Comparative Medicine, University of Virginia, Charlottesville, VA 22908;
Center for Blood Research, Harvard Medical School, Boston, MA 02115; and
¶ Department of Physiology, Louisiana State University Health Sciences Center, New Orleans, LA 70112
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
|
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Mice lacking E-, P-, and L-selectin (E-/-P-/-L-/-) have been generated by two groups. Robinson et al. (7) reported that the E-/-P-/-L-/- mice (Hynes mutation) developed ulcerative dermatitis although the frequency and age of onset were not evaluated. The E-/-P-/-L-/- mice described by Collins et al. (8) (Baylor mutation) are largely protected from ulcerative dermatitis seen in E-/-P-/- mice. Only 3 of 156 E-/-P-/-L-/- mice showed ulcerative dermatitis by 6 mo of age (8). However, this same line of E-/-P-/-L-/- mice (Baylor mutation) developed skin lesions at another facility (University of Washington, Seattle, WA), suggesting that lesion development is related to differences in animal husbandry and environment. Mice deficient in E- and P-selectin and ICAM-1 (E-/-P-/-I-/-) show a complete protection from the development of ulcerative dermatitis (9), suggesting a role for ICAM-1 in the development of ulcerative dermatitis that occurs in the absence of E- and P-selectin.
The relative protection from ulcerative dermatitis in E-/-P-/-L-/- and E-/-P-/-I-/- mice suggests that both L-selectin and ICAM-1 may serve a function critical to lesion development. L-selectin is necessary for lymphocyte homing into peripheral lymph nodes (10) and for lymphocyte recirculation (11). As a result, L-selectin-deficient mice have altered lymphocyte function, impaired T cell responses, and display very small lymph nodes (12). Because trafficking of naive lymphocytes is impaired in E-/-P-/-L-/- mice, the immune response to bacterial Ags present in the normal flora may be reduced. T cell-stimulating activity was also shown to be impaired in mice lacking ICAM-1, suggesting that ICAM-1 is an important costimulatory molecule (13). Based on these findings, we hypothesized that defective lymphocyte function in E-/-P-/-L-/- and E-/-P-/-I-/- mice protects these mice from lesion development. We inferred that the ulcerative dermatitis seen in E-/-P-/- mice may be lymphocyte-dependent. To test the hypothesis that the development of ulcerative dermatitis in E-/-P-/- mice requires mature T cells, we generated mice deficient in E- and P-selectin and the V(D)J recombination activation gene (Rag)3-1 (14) (E-/-P-/-Rag-1-/-) by cross-breeding E-/-P-/- with Rag-1-/- mice. Mice lacking the V(D)J Rag-1 do not have mature T or B lymphocytes (14). E-/- P-/-Rag-1-/- mice are healthier than E-/-P-/- mice and do not develop ulcerative dermatitis seen in E-/-P-/- mice.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Mice deficient in both E- and P-selectin and Rag-1 were generated by cross-breeding E-/-P-/- C57BL/6 N6 (1) and Rag-1-/- C57BL/6 N10 (The Jackson Laboratory, Bar Harbor, ME) mutants. F1 hybrids were back-crossed to E-/-P-/- to generate E+/-P+/-Rag-1+ and E-/-P-/-Rag-1+ mice. F1 hybrids were back-crossed to Rag-1-/- mice to generate E+/+P+/+Rag-1-/- and E+/-P+/-Rag-1-/-. Breedings were established from these offspring to generate all necessary control groups, including E+/+P+/+Rag-1+, E+/-P+/-Rag-1+, E+/+P+/+Rag-1-/-, E+/-P+/- Rag-1-/-, E-/-P-/-Rag-1+, and E-/-P-/-Rag-1-/- mice. Because there is no evidence for haplo-insufficiency in any of the selectin mutants, E+/-P+/- mice were pooled with E+/+P+/+ mice. Wild-type refers to E+/+P+/+Rag-1+ and E+/-P+/-Rag-1+ mice; Rag-1-/- refers to E+/+P+/+Rag-1-/- and E+/-P+/-Rag-1-/- mice; and E-/-P-/- refers to E-/-P-/-Rag-1+ mice.
Polymerase chain reaction
Genomic DNA was analyzed using PCR for the wild-type and knockout E- and P-selectin alleles. The P-selectin PCR assay used a forward primer from exon 3 of murine P-selectin (5'-TCA CGG GTG TTC TGT AGG AGG-3') and a reverse primer from exon 3 (5'-GGG GCC GAG TTA CTC TTG ATG-3'), yielding a 260-bp wild-type fragment. The P-selectin knockout allele was detected using a forward primer from exon 2 (5'-GCT GGC TGC CCA AAA GGT T-3') and a reverse primer from the inserted neomycin resistance gene (5'-ACC CGT GAT ATT GCT GAA GAG C-3'). This reaction yielded a 1300-bp mutant fragment. PCR conditions for the detection of the wild-type alleles were 94°C for 4 min, 35 cycles of 94°C for 30 s, 55°C for 30 s, and 72°C for 30 s, followed by 72°C for 7 min. PCR conditions for the detection of the P-selectin knockout alleles were 94°C for 4 min, 35 cycles of 94°C for 1 min, 61°C for 1 min, and 72°C for 1 min, followed by 72°C for 7 min.
The E-selectin PCR assay used a forward primer from exon 5 of murine E-selectin (5'-TTG GCT GTA AAA GGG GCT ACC-3') and a reverse primer from exon 6 (5'-CAT GAT GGC GTC TCG TTA TCC-3'), yielding a 1000-bp wild-type fragment. The E-selectin knockout allele was detected by a forward primer from hypoxanthine phosphoribosyltransferase gene (5'-ACT ATC AGT TCC CTT TGG GCG-3') and the reverse primer (5'-CAT GAT GGC GTC TCG TTA TCC-3'), which yields a 650-bp knockout fragment. PCR conditions for the detection of the E-selectin wild-type alleles were 94°C for 4 min, 35 cycles of 94°C for 1 min, 57°C for 1 min, and 72°C for 1 min, followed by 72°C for 7 min. PCR conditions for the detection of the E-selectin knockout allele were 94°C for 4 min, 35 cycles of 94°C for 1 min, 59°C for 1 min, and 72°C for 1 min, followed by 72°C for 7 min.
Serum G-CSF measurements
Serum G-CSF protein concentrations were determined using a specific enzyme-linked immunoassay using Ab pairs purchased from R&D Systems (Minneapolis, MN). For this purpose, 96-well plates (NuncImmunoplate Maxisorb; Nunc, Neptune, NJ) were coated with 2 µg/ml of capture Ab and incubated overnight at 4°C. The plates were then washed five times with wash buffer (0.05% Tween 20 in PBS) and blocked with 200 µl 2% BSA in wash buffer for 2 h at room temperature. G-CSF standards and samples were diluted in wash buffer containing 2% FCS. Both standards (31.25–1000 pg/ml) and samples (50 µl) were added to wells and the plates were incubated for 1 h at 37°C. After washing, 50 µl of biotinylated anti-G-CSF (0.1 µg/ml in dilution buffer) was added and the plates were incubated for 1 h at 37°C. Wells were then washed and incubated for 1 h after adding 100 µl of 0.1 µg/ml peroxidase-conjugated streptavidin (Jackson ImmunoResearch Laboratories, West Grove, PA) in dilution buffer. After washing the plate, 100 µl of tetramethylbenzidine (Sigma-Aldrich, St. Louis, MO) was added as substrate and color was allowed to develop for 30 min in the dark. After stopping the reaction with 50 µl of 3 M H2SO4, ODs were determined at 450 nm. G-CSF concentrations were calculated from the standard curve using log-log linear regression.
Flow cytometry
Peripheral blood was collected from the tail vein. Expression of CD4 and CD8 on peripheral lymphocytes was determined by direct immunofluorescence to identify mice lacking the V(D)J Rag-1. Lymphocytes were identified and gated by forward and side scatter properties. CD8+ cells were identified by FITC-labeled mAb 53-6.7 (BD PharMingen, San Diego, CA) staining. CD4+ cells were identified by FITC-labeled mAb GK1.5 (BD PharMingen) staining. Cells were analyzed by forward scatter, side scatter, and FITC fluorescence using a laser flow cytometer (FACSCan; BD Biosciences, Mountain View, CA). Data are presented as fluorescence histograms of CD4 and CD8 expression on a four-decade log scale.
Histopathology
Age-matched wild-type, Rag-1-/-, E-/-P-/- with and without ulcerative dermatitis, and E-/-P-/-Rag-1-/- mice were euthanized by cervical dislocation under anesthesia for histopathological analysis. After gross examination for any discernable pathology, lung, intestine, kidney, cervical lymph nodes, liver, spleen, and skin samples were fixed in 10% buffered formalin (Sigma-Aldrich), embedded in paraffin, sectioned, and examined after H&E staining.
Microbiological analyses
Tissue samples for microbiologic analysis were obtained under sterile conditions from the lung, liver, and spleen from wild-type Rag-1-/-, E-/-P-/- with and without ulcerative dermatitis, and E-/-P-/-Rag-1-/- mice. Tissues were homogenized with a sterile grinder in 0.5 ml PBS. A 100-µl sample from each tissue was plated onto trypticase soy agar with 5% sheep RBCs (Difco, Detroit, MI). Plates were observed at 24 and 48 h for growth.
Statistics
Total leukocyte counts, neutrophil counts, mononuclear cell counts, and serum G-CSF levels between groups were compared using Kruskal-Wallis One-Way ANOVA on ranks and pairwise multiple comparison by Dunn’s method. Spleen weights and lymph node weights between groups were compared using Kruskal-Wallis one-way ANOVA and pairwise multiple comparison by the Tukey test. Statistical significance was set at p < 0.05.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Mice deficient in E- and P-selectin and Rag-1 were generated by
cross-breeding E-/-P-/-
C57BL/6 N6 (1) and Rag-1-/-
C57BL/6 N10 (The Jackson Laboratory) mutants. This resulted in all
necessary control groups, including
E+/+P+/+Rag-1+
(wild-type),
E+/-P+/-Rag-1-/-
(Rag-1-/-),
E-/-P-/-Rag-1+/+
(E-/-P-/-), and
E-/-P-/-Rag-1-/-
(Fig. 1
A; Table I). Mice
expressing E- and P-selectin
(E+/+P+/+), heterozygous
for E- and P-selectin
(E+/-P+/-), and
homozygous for a null mutation in E- and P-selectin
(E-/-P-/-) were
determined by PCR. Consistent with previous results (1, 4)
and the close distance of the E- and P-selectin genes, we found that E-
and P-selectin cosegregated in all mice tested.
|
|
A, lane 1).
No band is detectable from either of the knockout reactions in
E+/+P+/+ mice (Fig. 1A, lane 2). A heterozygous mutation for E- and
P-selectin (E+/-P+/-)
results in both wild-type and knockout bands from the P-selectin and
the E-selectin reactions (Fig. 1A, lanes 3 and
4). Mice that have a null mutation in E- and P-selectin
(E-/-P-/-) show no
band from either wild-type reaction (Fig. 1A, lanes
5 and 7) and a single band from the respective knockout
reactions (Fig. 1A, lanes 6 and
8). Verification for a null mutation for Rag-1 was
determined by flow cytometry. Mice deficient in Rag-1 have no
CD4+ or CD8+ lymphocytes in
blood (Fig. 1B, bottom panels). White blood cell counts and serum G-CSF levels
Because mice deficient in Rag-1 do not develop functional T or B
lymphocytes, Rag-1-/- mice show a significant
reduction of mononuclear cells in the peripheral circulation compared
with wild-type mice (Table II).
Rag1-deficient mice had neutrophil counts similar to wild-type mice
(Table II). E-/-P-/-
mice displayed significantly elevated leukocyte counts compared with
wild-type and Rag-1-/- mice.
E-/-P-/- mice were
severely neutrophilic (Table II). Neutrophil counts in
E-/-P-/-Rag-1-/-
mice were almost as high as in
E-/-P-/- mice. However,
E-/-
P-/-Rag-1-/- mice
had significantly fewer circulating mononuclear cells than
E-/-P-/- mice
(Table II).
|
).
E-/-P-/-Rag-1-/-
mice showed G-CSF levels similar to
E-/-P-/- mice (Table II). Similar increases in neutrophil counts and G-CSF levels in
E-/-P-/-Rag-1-/-
mice compared with
E-/-P-/- mice show that
the level of neutrophilia in these mice is not dependent on lymphocyte
function (Table II). Clinical and pathological findings
E-/-P-/-Rag-1-/-
mice were generally healthier than
E-/-P-/- mice.
E-/-P-/-Rag-1-/-
breeders consistently produced litters for 
1 year.
E-/-P-/- breeders only
occasionally generated litters until 5–6 mo of age in the same
specific pathogen-free barrier facility. Therefore, the
E-/-P-/- colony was
maintained with heterozygous breeders.
Approximately 50% (18 of 35) of the
E-/-P-/- mice 6–12 mo
old developed conjunctivitis and/or facial and submandibular ulcerative
dermatitis (Fig. 2C).
Initially, these mice showed hair loss around the eyes and the
development of conjunctivitis. This was followed by hair loss on the
nose and neck and subsequent skin lesion development (Fig. 2A). These lesions have primarily been shown to be colonized
by commensal bacteria, but not specific pathogens (1, 4).
E-/-P-/-Rag-1-/-
mice were completely protected from the development of ulcerative
dermatitis (Fig. 2B). None of the 62
E-/-P-/-Rag-1-/-
mice developed facial or submandibular ulcerative dermatitis, including
25 mice older than 12 mo (Fig. 2C).
|
To investigate possible bacteremia and colonization of internal organs by commensal bacteria in E-/-P-/- mice, liver, lung, and spleen samples were analyzed for bacterial growth. No positive cultures were obtained in wild-type (n = 4), Rag-1-/- (n = 5), E-/-P-/- showing ulcerative dermatitis (n = 5), E-/-P-/- with no signs of ulcerative dermatitis (n = 5), or E-/-P-/-Rag-1-/- (n = 8) mice.
Histopathology
Cervical skin from
E-/-P-/-Rag-1-/-
mice showed an intact epidermis, no dermal infiltrate, and no bacterial
colonization (Fig. 3C). Tissue
samples from E-/-P-/-
mice with ulcerative skin lesions showed areas of epidermal loss as
well as hyperplastic epithelium, and a dermal infiltrate consisting of
mixed inflammatory cells, including mast cells,
macrophages, and mononuclear cells (Fig. 3, A and
B). Areas of ulceration also contained large numbers of
bacterial colonies both on the denuded surface and infiltrating the
underlying dermis (Fig. 3B). Neutrophils were noticeably
lacking despite extensive bacterial colonization. There was also
evidence of loss of sebaceous glands and distortion of hair
follicles.
|
).
As expected, the lymphoid follicles (white pulp) were absent in
Rag-1-/- mice (Fig. 4), but some myeloid and
erythroid precursor cells were present.
E-/-P-/- mice showed a
profound expansion of red pulp due to extramedullary hemopoiesis,
disrupting the normal splenic architecture (Fig. 4). Large numbers of
megakaryocytes, erythroid, myeloid precursors, neutrophils, and plasma
cells were present throughout the spleen. The white pulp in these mice
was smaller, distorted, and less distinct (Fig. 4).
E-/-P-/-Rag-1-/-
mice exhibited a similar pattern of extramedullary hemopoiesis;
however, these mice showed an absence of white pulp (Fig. 4). As a
result of extramedullary hemopoiesis, the spleen weights normalized for
body weights in
E-/-P-/- and
E-/-P-/-Rag-1-/-
mice were increased 3- and 2-fold, respectively, compared with
wild-type and Rag-1-/- mice (Table III).
|
|
) were indistinguishable from wild-type
mice (data not shown). Mice lacking E- and P-selectin showed degrees of
lung pathology varying from mild to severe. Interstitial cellularity
was increased secondary to leukocyte infiltration (Fig. 5), and in more
severe cases evidence of interstitial edema and hemorrhage were
present.
E-/-P-/-Rag-1-/-
mice generally had less inflammatory cell infiltrate in the lungs
compared with E-/-P-/-
mice (Fig. 5). Where seen, infiltrates appeared to be more focal than
diffusely distributed.
|
) compared with wild-type mice
(11.0 ± 0.1 mg; Fig. 6; Table III).
E-/-P-/- mice displayed
cervical lymphadenopathy (19.6 ± 2.4 mg), the normal
corticomedullary architecture was disrupted, and lymph nodes were
infiltrated by inflammatory mononuclear and plasma cells (Fig. 6). In
some cases, particularly in mice with severe skin lesions overlying the
submandibular lymph nodes, lymphoid follicles were completely effaced
by plasma cells.
E-/-P-/-Rag-1-/-
mice had very small lymph nodes that were not different from
Rag-1-deficient mice (3.7 ± 0.7 mg; Table III). These lymph nodes
contained few lymphocytes, no plasma cell infiltrate, and more
connective tissue and fibroblasts (Fig. 6). Colon, small intestine,
liver, and kidney from all groups showed no significant pathology.
|
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
E-/-P-/- mice develop ulcerative dermatitis of the nose and cervical regions at all institutions where they are bred. Mice lacking all three selectins (E-/-P-/-L-/-) develop ulcerative dermatitis in some facilities (University of Washington, Seattle, WA; and Harvard Medical School, Boston, MA), while they are largely protected from disease development in other facilities (Baylor College of Medicine, Houston, TX; University of Virginia, Charlottesville, VA). The causative trigger of ulcerative dermatitis is not known but appears to be related to environmental and/or housing conditions. We hypothesized that the skin pathology results from a compensatory immune response to commensal bacteria, including plasma cell hyperproliferation, and that a diminished immune response in the absence of ICAM-1 (E-/-P-/-I-/-) and L-selectin (E-/-P-/-L-/-) protects these mice from disease development. In this study, we show that an absence of functional T and B cells (E-/-P-/-Rag-1-/-) completely protects these mice from the development of ulcerative dermatitis, although housed in the same environment.
Histological analysis of skin sections of
E-/-P-/- mice reveals a complete absence of
neutrophils in skin lesions, showing that neutrophils are not recruited
to these areas of infection in the absence of E- and P-selectin.
Interestingly, significant levels of lymphocyte recruitment occur
despite the lack of E- and P-selectin. Lymphocyte recruitment to skin
lesions is likely due to the critical involvement of
4 integrins in the recruitment of lymphocytes
to sites of chronic inflammation. We have shown that lymphocyte
recruitment and adhesion were not affected in chronic models of
inflammation (thioglycolate-induced peritonitis and 6-h TNF--induced
inflammation of the mouse cremaster muscle, respectively) in mice
lacking E-, P-, and L-selectin and ICAM-1
(E-/-P-/-L-/-I-/-;
Ref. 15). Leukocyte adhesion was almost completely
inhibited only with anti-4 integrin mAb
pretreatment. T lymphocyte migration to dermal inflammation induced by
IFN-
, TNF-, LPS, or poly inosine-cytosine was markedly reduced by
anti-4 integrin treatment (50–60%). T
lymphocyte migration was inhibited by only 30–60% with anti-E-
and P-selectin treatment compared with 10–40% with either anti-E-
or P-selectin treatment. Blockade of both E- and P-selectin and
4 integrin function severely impaired
lymphocyte recruitment in these models of dermal inflammation (>90%;
Ref. 16). Although E- and P-selectin have a role in
lymphocyte recruitment, they are not strictly required in some models
of inflammation.
The molecular mechanisms linking a dysregulated immune response to
conjunctivitis and cervical ulcerative dermatitis remain unknown
and were not addressed in this study. Decreased neutrophil trafficking
in E-/-P-/- mice may
result in expansion of commensal bacteria and an increased antigenic
load which may lead to the release of proinflammatory cytokines which
potentiate host tissue damage and ulceration. Others have shown that T
cells are actively involved in inflammatory skin disorders, including
allergic contact dermatitis, atopic dermatitis, and psoriasis
(17, 18, 19). Cytokines released by activated skin
infiltrating T lymphocytes, such as TNF-, IFN-, and IL-4, induce
the activation of keratinocytes to produce proinflammatory chemokines,
including inducible protein-10, IFN-inducible T cell chemoattractant,
and monokine induced by IFN-, which results in the continued
recruitment of CXCR3+ T cells (17).
Chronic abnormal chemokine and proinflammatory cytokine production from
activated infiltrating T cells and keratinocytes may lead to host
tissue damage and lesion development. Consistent with this
interpretation, removing lymphocyte function in
E-/-P-/-Rag-1-/-
mice eliminates this overshooting response to normal flora Ags and
protects these mice from tissue damage and ulceration despite defective
neutrophil trafficking. Remarkably, the same commensal bacteria
colonizing dermal lesions in
E-/-P-/- mice do not
invade
E-/-P-/-Rag-1-/-
mice. This suggests that the dysregulated immune response in
E-/-P-/- mice is
pathogenic rather than protective under vivarium conditions.
Secretion of various cytokines from T cells promotes neutrophil
recruitment and stimulates granulopoiesis, linking the acquired immune
system with the innate system. IL-17 released from activated T cells
has been shown to stimulate neutrophil recruitment through the release
of keratinocyte-derived chemokine and macrophage-inflammatory
protein-2 (20, 21). T cells regulate hemopoiesis through
secretion of various cytokines, including IL-2, IL-3, IL-6, IL-8,
IL-17, and GM-CSF (22, 23). IL-17 stimulates
granulopoiesis in vivo through G-CSF release (22).
E-/-P-/-Rag-1-/-
mice show elevated serum G-CSF and levels of neutrophilia similar to
E-/-P-/- mice,
suggesting that lymphocyte-independent mechanisms completely regulate
the elevated neutrophil levels seen in these mice (20,000
polymorphonuclear cells/µl). Surprisingly,
E-/-P-/-Rag-1-/-
mice do not show neutrophil levels higher than
E-/-P-/- mice. Although
granulopoiesis can be enhanced or modulated by the acquired immune
system (22), lymphocyte-independent granulopoiesis in
E-/-P-/-Rag-1-/-
mice is sufficient to compensate for the loss of the acquired immune
system. This suggests there is an independence of the acquired and
innate immune systems in regulating this level of neutrophilia.
In conclusion, we show that E-/-P-/-Rag-1-/- mice which lack functional lymphocytes display a healthier phenotype than E-/-P-/- mice, although circulating neutrophil counts and serum G-CSF levels are similar to E-/-P-/- mice. E-/-P-/-Rag-1-/- mice did not have hypercellular cervical lymph nodes and showed reduced leukocyte infiltrate in the lungs. Most importantly, E-/- P-/-Rag-1-/- mice were completely protected from the ulcerative dermatitis that develops in E-/-P-/- mice. We conclude that a dysfunctional immune response in E-/-P-/- mice resulting from impaired leukocyte trafficking leads to ulcerative dermatitis in E-/-P-/- mice.
|
Acknowledgments |
|---|
|
Footnotes |
|---|
2 Address correspondence and reprint requests to Dr. Klaus Ley, Cardiovascular Research Center, University of Virginia, Health System Box 801394, Charlottesville, VA 22908. E-mail address: klausley{at}virginia.edu
3 Abbreviations used in this paper: Rag, recombination activation gene.
Received for publication June 4, 2002. Accepted for publication August 29, 2002.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
) and type 2 (IL-4, IL-5) cytokines in T-helper and T-suppressor/cytotoxic cells in rheumatoid arthritis, allergic asthma and atopic dermatitis. Cytokine 11:783.[Medline]
chemokine from mesothelial cells. J. Immunol. 165:5814.This article has been cited by other articles:
|
|
 |
|
  E. Smith, S. von Vietinghoff, M. A. Stark, A. Zarbock, J. M. Sanders, A. Duley, J. Rivera-Nieves, T. P. Bender, and K. Ley T-Lineage Cells Require the Thymus but Not V(D)J Recombination to Produce IL-17A and Regulate Granulopoiesis In Vivo J. Immunol., November 1, 2009; 183(9): 5685 - 5693. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Urzainqui, G. Martinez del Hoyo, A. Lamana, H. de la Fuente, O. Barreiro, I. M. Olazabal, P. Martin, M. K. Wild, D. Vestweber, R. Gonzalez-Amaro, et al. Functional Role of P-Selectin Glycoprotein Ligand 1/P-Selectin Interaction in the Generation of Tolerogenic Dendritic Cells J. Immunol., December 1, 2007; 179(11): 7457 - 7465. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Zaph and P. Scott Th1 Cell-Mediated Resistance to Cutaneous Infection with Leishmania major Is Independent of P- and E-Selectins J. Immunol., November 1, 2003; 171(9): 4726 - 4732. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
